Closed Kit for Tissue Containment and Stabilization for Molecular and Histopathology Diagnostics

ABSTRACT

A container for storing a biological sample is disclosed. The container includes a housing having a closed end, an open end, and a sidewall extending therebetween defining a container interior. The container has a removable closure for enclosing the open end of the housing and a sample holder for housing a biological sample detachably connected to the closure and insertable within the container interior. A port is disposed within the closure adjacent the sample holder to allow fluid to pass therethrough into the container interior. An injection device for engaging the port may also be provided. A first fluid may be initially provided within the container interior and a second fluid may be subsequently injected by the injection device through the port into the container interior.

CROSS REFERENCE TO RELATED APPLICATION

This application claims priority to U.S. Provisional Patent ApplicationNo. 60/982,049, filed Oct. 23, 2007, entitled “Closed Kit for TissueContainment and Stabilization for Molecular and HistopathologyDiagnostics”, the entire disclosure of which is herein incorporated byreference.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to a closed kit system for storing abiological sample. More particularly, the present invention relates to aclosed kit system for storing a biological tissue specimen in a reagentor, if so desired, in multiple reagents for molecular or diagnostictesting and/or histological testing.

2. Description of Related Art

Biological samples are often obtained by a researcher or clinician fordiagnostic evaluation to determine the presence of certain diseases andto determine an appropriate treatment for the disease. Tissue samplesare often obtained from a patient for molecular diagnostic and nucleicacid analysis, particularly RNA and DNA analysis, which have becomecommon place in research for the treatment of numerous diseases. Anessential requirement for accurate RNA and DNA analysis is the presenceof high quality and intact RNA and DNA within the biological sample.

Oftentimes, the histologic or cytologic analysis will be performedimmediately after the sample is removed from the patient or source toavoid molecular changes that may occur during storage. These changes,such as gene transcription, result from the degradation of the nucleicacids within the sample caused by exposure of an untreated sample tocertain environmental stresses. However, analysis of the sampleimmediately after the sample is collected is often impossible orimpractical. Therefore, it is necessary to provide a system for storinga sample under controlled conditions for a certain period of time whilemaintaining the structural and molecular integrity of the sample.

Traditionally, one way of accomplishing this storage is by submergingthe sample in a single fixative reagent. A typical fixative reagent isten percent (10%) formalin but may also include water, misciblealcohols, ethanol/acetone mixtures, and ethanol/acetic acid mixtures.The containers used for such storage are generally composed of a singleintegral cavity which could house an effective volume of reagent totreat a particular biological tissue sample. The biological tissuesample is placed in the container along with the reagent, the containeris closed, and the sample is then stored and transported while beingpreserved by the fixative agent. An example of such a container can beseen in U.S. Pat. No. 7,147,826 to Haywood et al. Such containers haveexperienced some success in the industry, but are subject to certainlimitations.

SUMMARY OF THE INVENTION

In one embodiment of the present invention a system for storing abiological sample includes a container having a closed end, an open end,and a sidewall extending therebetween defining a container interior. Thecontainer also includes a removable closure for enclosing the open endof the container, and a sample holder for housing a biological sample.The sample holder is insertable within the container interior. A port isdisposed within one of the container and the removable closure, and aninjection device is engageable with the port to allow a fluid to passfrom within the injection device into the container interior.

The sample holder may be detachably connected to the removable closure.In one configuration, the port is disposed within the closure adjacentthe sample holder. Optionally, the system includes a platform attachedto the removable closure and adapted for receiving the sample holder.The sample holder may be positioned in fluid-access with the port suchthat fluid introduced through the port passes directly through thesample holder. In a further configuration, the sample holder is ahistology cassette.

The system may also include a vent adapted to discharge a volume of airfrom the container interior. The vent may be disposed within theremovable closure. Optionally, the vent transitions from a first closedposition to a second open position when the volumetric pressure withinthe container interior reaches a threshold value. The removable closureand the open end of the container may be threadably matable.

In a further configuration, the injection device is a syringe.Optionally, the injection device may engage the port and, uponengagement, the port may transition from a first closed position to asecond open position. A first fluid may be disposed within the containerinterior, and a second fluid may be disposed within the injectiondevice, with the second fluid being different than the first fluid. Inanother configuration, the port may include a luer-activated valve.

In another embodiment of the present invention, a container for storinga biological sample includes a housing having a closed end, an open end,and a sidewall extending therebetween, defining a container interior.The housing also includes a removable closure for enclosing the open endof the housing, and a sample holder for housing a biological sample. Thesample holder may be detachably connected to the removable closure, andinsertable within the container interior. A port is disposed within theremovable closure adjacent the sample holder. The port is adapted toallow a fluid to pass therethrough from a location exterior to thecontainer to the container interior.

Optionally, the sample holder is a histology cassette. The container mayalso include a vent disposed within the removable closure adapted todischarge a volume of air from the container interior. A first fluid maybe disposed within the container interior, and the port may be adaptedto receive an injection device at least partially therein, fortransferring a second fluid into the container interior, the secondfluid different than the first fluid.

In yet another embodiment of the present invention, a method of storinga biological sample includes the step of providing a container having ahousing having a closed end, an open end, and a sidewall extendingtherebetween defining a container interior. The container interior has afirst fluid disposed therein. The container also includes a removableclosure for enclosing the open end of the housing, and a sample holderhousing a biological sample. The sample holder is detachably connectedto the removable closure and is disposed within the first fluid of thecontainer interior. The container also includes a port disposed withinthe removable closure adjacent the sample holder. The method includesthe step of engaging an injection device containing a second fluid withthe port, with the second fluid different than the first fluid. Themethod also includes the step of injecting the second fluid from theinjection device through the port into the container interior.

Further details and advantages of the invention will become apparentupon reading the following detailed description in conjunction with theaccompanying figures, wherein like parts are designated with likereference numerals throughout.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a perspective view of a closed kit system pursuant to oneembodiment of the present invention.

FIG. 2 is a perspective view of a container of the kit of FIG. 1 shownwithout a closure.

FIG. 3 is a perspective view of the container of the kit of FIG. 1 shownwith the closure according to one embodiment.

FIG. 4 is a cross-sectional view of the container shown in FIG. 3 takenacross line A-A.

FIG. 5 is a perspective view of the closure of the kit of FIG. 1oriented so the bottom surface faces upward showing a sample holderconnected to the bottom surface.

FIG. 6 is a side view of an injection device according to oneembodiment.

FIGS. 7A and 7B are perspective views of the sample holder according toone embodiment shown in the open and closed position, respectively.

FIG. 8 is a perspective view of a closed kit system as in FIG. 1 shownwith a different embodiment of the injection device.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS

For purposes of the description hereinafter, spatial orientation terms,if used, shall relate to the referenced embodiment as it is oriented inthe accompanying drawing figures or otherwise described in the followingdetailed description. However, it is to be understood that theembodiments described hereinafter may assume many alternative variationsand embodiments. It is also to be understood that the specific devicesillustrated in the accompanying drawing figures and described herein aresimply exemplary and should not be considered as limiting.

The closed kit system of the present invention allows for storage of abiological sample, such as a tissue sample for molecular and histologydiagnostics, and in particular histopathology testing. Particularly, thesystem includes a container with an open end, a closed end, and asidewall extending between the open end and the closed end defining acontainer interior. The system further includes an injection device anda port that allows fluid to pass from the injection device to thecontainer interior. Accordingly, the injection device is capable ofinjecting a liquid medium into the container. In this manner, a tissuesample contained in the container may be handled or processed prior tocontacting the tissue with the solution in the injection device. As willbe discussed in greater detail herein, in one embodiment of theinvention, the container interior does not contain a solution andrepresents a storage chamber, and the injection device includes a liquidmedium, such as a reagent in the form of a tissue fixative solution forfixing a sample for histopathology diagnostics. In this manner, a tissuesample may be placed within the empty container interior, and whendesired, the injection device may be used to introduce a reagent intothe container interior so as to place the tissue sample in fluid contactwith the reagent within the injection device.

In a further embodiment of the invention, the container interior maycontain a first fluid or reagent, such as a tissue fixative solution,and the injection device may contain a second fluid or reagent, such asa nucleic acid stabilization solution, such that a tissue sample may beplaced in the container interior in fluid contact with the first fluidfor a desired time period, after which time the injection device may beused to inject a second fluid into the container interior so as to placethe tissue sample in fluid contact with the second fluid. Theembodiments described herein are representative of closed kit systemscapable of use in any of these manners.

Referring to the drawings in which like reference characters refer tothe like parts throughout the several views thereof, FIG. 1 generallydepicts a closed kit system, such as system 10, for the storage of abiological tissue sample. System 10 generally includes a container 20, asample holder 50, an injection device 110, and a port 76 which receivesinjection device 110 and allows fluid to pass from injection device 110into container 20.

Container 20 includes a closed end 28, an open end 24, and a sidewall 32extending between open end 24 and closed end 28 defining a containerinterior 36. In one embodiment, as depicted in FIG. 1, container 20 iscylindrical in shape, although it is contemplated that container 20 canbe formed as any shape consistent with the intended use of thedescriptions herein. Container 20 may be constructed of any knownmaterial, such as glass or plastic, and is desirably molded of apolymeric material.

Sample holder 50 is further provided for use with system 10, and isadapted to be received within the interior 36 of container 20. Sampleholder 50 may be in the form of a conventional histology cassette (a“histo-cassette”) as is known in the art for storing a biological tissuesample during preparation of the sample for diagnostic testing. Suchsample holders or histo-cassettes are known for containing biologicalspecimens during processing with fluids to prepare the specimen forlater analyses. Typically, such sample holders or histo-cassettes aregenerally rectangular, planar housing structures having an internalcavity, with a plurality of openings through the wall surface to providefluid flow through the housing. Often, a removable or openable coverencloses the structure, such as through a hinge situated along one endof the housing structure for providing a door-like cover to the housingstructure. Also, a planar surface, which may be slanted, is oftenprovided in such sample holders or histo-cassettes acting as a surfacefor labeling or writing. The dimensions for such a sample holder, forexample, may include a height of about 0.3 inch (plus or minus 0.1inch), a length of about 1.73 inches (plus or minus 0.1 inch), and awidth of about 1.12 inches (plus or minus 0.1 inch). Examples of sampleholders that may be useful herein are shown in U.S. Pat. No. 4,220,252to Beall et al. and U.S. Pat. No. 4,034,884 to White, both of which areexpressly incorporated herein by reference.

For example, as shown in FIGS. 7A and 7B, sample holder 50 includes agenerally rectangular planar housing 52 having opposing walls definingan internal cavity 54 for holding a biological tissue sample therein. Atleast one of the walls of housing 52 may be slanted, such as slantedwall 55, providing a surface for applying a label or for writing, so asto provide a mechanism for identification of a sample contained withinsample holder 50 as appropriate. Housing 52 of sample holder 50 is aclosable structure, and may include a hinged door-like structure 58attached with housing 52 thereby permitting access to the internalcavity 54 for storing a tissue sample within or removing a tissue samplefrom internal cavity 54. The door-like structure 58 may be integrallyformed with housing 52 so as to provide a unitary structure with thedoor 58 connected to housing 52 through a flap to provide a mechanismfor pivoting door 58 with respect to housing 52, or door 58 may beotherwise connectable to housing 52, such as through a pivot point 53acting as a hinge for opening door 58 from one side of housing 52 togain access to the internal cavity 54. Housing 52 of sample holder 50includes at least one, and preferably a plurality of fluid openings 56adapted to allow fluid to flow therethrough. In this manner, whenhousing 52 is positioned within first chamber 20, fluid within firstchamber 20 can flow through openings 56 and contact the biologicaltissue sample contained within internal cavity 44.

System 10 also includes a cover or closure 60 for enclosing open end 24.Closure 60 may consist of a removable or pierceable membrane, such as awax or polymer material, that is affixed to container 20 and enclosesopen end 24. In one embodiment, shown in FIG. 3, closure 60 isrepresented as a cap or lid, desirably constructed of a molded polymermaterial. Closure 60 may be threadably matable with container 20 by wayof first threaded members 64 disposed within closure 60 andcorresponding second threaded members 40 disposed on the exterior ofcontainer sidewall 32 near open end 24. However, closure 60 can engagecontainer 20 in any manner, such as a frictional fit, interlockingstructural engagement, or other manner providing a liquid tight seal.

System 10 optionally includes structure for mating sample holder 50 withclosure 60. When sample holder 50 is mated with closure 60, sampleholder 50 will enter container interior 36 when closure 60 engagescontainer 20 as previously described. In one embodiment, mating isaccomplished by providing sample holder 50 as an integral part connectedto or formed with closure 60. Alternatively, sample holder 50 can bemated with closure 60 through a receiving member 68 extending from thebottom surface of closure 60 as shown in FIGS. 4-5. Receiving member 68is capable of, detachably or permanently, securing sample holder 50 toclosure 60 through, for example, a snap-fit engagement. In theembodiment shown in FIG. 5, receiving member 68 includes a platform 66and a series of fingers 70 extending from a bottom surface of closure 60for accommodating sample holder 50 therein. Platform 66 may includestructure for maintaining sample holder 50 attached to closure 60 asshown in FIG. 5, such as in a snap-fit engagement and sample holder 50may be releasable from platform 66. In particular, platform 66 may be agenerally rectangular structure defining a rectangular recess foraccommodating the general size and shape of sample holder 50. Platform66 may include one or more fingers 70 extending therefrom for engagingwith sample holder 50, thereby maintaining sample holder 50 within therecess or cavity 72 defined by platform 66. Such fingers 70 may bedeflectable, such that when an edge of sample holder 50 adjacent theslanted wall 55 is held in place against a corresponding protrusion orfinger (not shown) of the platform 66, and sample holder 50 is pushedinto the recess of platform 66, fingers 70 deflect away from the wallsof sample holder 50 and then return to their initial position, therebysnapping sample holder 50 in place. Fingers 70 may lock sample holder 50in place permanently with respect to platform 66 and closure 60, or maybe deflectable so as to remove sample holder 50 from platform 66 ifdesired.

Platform 66 may also be provided with a general shape so as to permitopening of door 58 of sample holder 50 while maintaining housing 52 ofsample holder 50 contained therein, thereby providing access to theinterior cavity 54 of sample holder 50 while sample holder 50 is held inplace within platform 66 and with respect to closure 60.

In this arrangement, cavity 72 may be created between platform 66, thebottom surface of closure 60, and fingers 70. Fluid contained incontainer interior 36 may flow throughout cavity 72. Optionally,platform 66 contains a plurality of openings thereon allowing fluidwithin cavity 72 to flow through platform 66 and contact sample holder50 and the biological sample contained therein. In one embodiment, whenport 76 is integrated into closure 60, as discussed below, platform 66can be positioned so that it is aligned with port 76, shown in FIG. 5.In this arrangement, when a fluid passes through port 76, the fluidquickly comes into contact with platform 66 and then sample holder 50via the plurality of openings on platform 66.

System 10 also includes a port 76, which allows fluid to pass frominjection device 110 to container interior 36 when port 76 is engaged byinjection device 110 or, more particularly, a nozzle 114 thereof.Generally, port 76 is a valve normally in the closed position that cantransition to an opened position upon insertion of nozzle 114 into anend of port 76 adapted to receive nozzle 114. When in the open position,port 76 allows fluid to flow through it in at least one direction, andport 76 is able to resist large back pressures when in the closedposition. Any type of valve capable of allowing fluid to flow in atleast one direction currently known or hereinafter discovered may beused as port 76. In one embodiment, port 76 is comprised of aluer-activated valve, which is commonly known in the industry. Examplesof valves that can be used as port 76 within system 10 are shown in U.S.Pat. No. 5,775,671 to Cote, Sr. and U.S. Pat. No. 7,100,890 to Cote Sr.et al., both of which are expressly incorporated herein by reference.

In one embodiment, shown in FIGS. 3-4, port 76 is embedded, orintegrated, into closure 60. In this embodiment, port 76 is comprised oftwo ends where one end is located adjacent to one surface of closure 60and the other end is located adjacent to the other surface of closure60. In this arrangement, port 76 forms a passageway through closure 60allowing fluid to flow through closure 60 and into the containerinterior 36. In another embodiment, port 76 is embedded, or integrated,into sidewall 32 of container 20 and forms a passageway through whichfluid flows through sidewall 32 before entering container interior 36.In a further embodiment, the port 76 is positioned adjacent the sampleholder 50 to ensure sufficient wetting of a sample contained within thesample holder 50 by fluid injected from the injection device 110. In oneconfiguration, the port 76 is disposed within the closure 60 directlyabove the sample holder 50, such that when fluid is injected from theinjection device 110 through the port 76, the fluid directly contactsthe sample disposed within the sample holder 50. Optionally, theplatform 66 for holding the sample holder 50 may be positioned influid-access with the port 76 such that fluid introduced through theport 76 passes directly through the platform 66 and/or sample holder 50.The port 76 is adapted to allow a fluid to pass therethrough from alocation exterior to the housing 52 and closure 60, to the containerinterior 36. The sample holder 50 may be positioned directly under theport 76, or provided in fluid communication with the port 76 through aseries of baffles and/or channels for directed fluid therethrough.

The system 10 also includes an injection device 110, which is used toinject a reagent or solution into container interior 36 where it cancontact a tissue sample that has been placed within container interior36. Injection device 110 is received by port 76 and, once so received,is capable of injecting a fluid through the passageway created by port76 into container interior 36. In one variation, injection device 110includes a nozzle 114 which engages port 76. Optionally, nozzle 114 is aluer fitting. In one embodiment, injection device 110 may contain a setamount of solution or reagent within it, such as where injection device110 is a syringe or a blister package, both of which are commonly knownin the industry. In another embodiment, injection device 110 isconnected to a reservoir or tank by, for example, a hose or tube. Thereservoir contains the reagent or solution that is to be injected intocontainer interior 36. In this embodiment, once injection device 110 isengaged with port 76, an amount of the solution can flow from thereservoir through injection device 110 and into container interior 36,where it can contact the tissue sample therein.

System 10 also may include a vent 88 for removing air or other mediafrom container interior 36 that is displaced during the injectionprocess. Vent 88 acts as a transitional barrier between containerinterior 36 and the external environment and is transitional from afirst closed position where no media is released from container interior36 to a second open position where media, preferably air, can flow fromcontainer interior 36 to the surrounding environment. Vent 88 can be anytype of valve or stopcock currently known or hereinafter discovered.Vent 88 can operate to automatically release air from container interior36 when the volumetric pressure within container interior 36 reaches athreshold value. Alternatively, vent 88 can be manually transitionedfrom the closed position to the open position. In one embodiment, vent88 is embedded, and integrated, into closure 60 so that vent 88, when inthe open position, forms a passageway through closure 60 into containerinterior 36. In another embodiment, vent 88 is embedded, and integrated,into sidewall 32 of container 20 thus creating a passageway throughsidewall 32 to container interior 36 when in the open position.

System 10 may be assembled and provided with liquid media, such assolutions or reagents, stored within container interior 36 and/orinjection device 110 at the point of manufacture. Alternatively, anysuch liquid media may be filled into container interior 36 and/orinjection device 110 at any point prior to use, such as directly priorto inserting a tissue sample into sample holder 50.

System 10 may be provided for use with a one reagent scheme. In thismanner, a single reagent solution, such as a tissue fixative likeformalin, may be provided within injection device 110. Such fixativesolutions stabilize the RNA within a tissue sample for conductingmolecular diagnostic testing. Alternatively, system 10 may be providedfor use with a two solution or a two reagent scheme. For example, a washsolution may be provided in injection device 110 so as to dilute thefirst reagent fixative in container interior 36. It is also possiblethat container interior 36 and injection device 110 each contain thesame reagent since it may be advantageous to refresh the reagent incontainer interior 36 with additional reagent after a period of time haspassed. Or, a first reagent solution, such as a tissue fixative likeformalin, may be used within container interior 36, and a second reagentsolution, such as a stabilizer in the form of a nucleic acidstabilization reagent for stabilizing the morphology of the tissuesample, may be provided within injection device 110.

Any reagents may be used with the container of the present invention.For example, the fixative may be formalin, ethanol solutions, Carnoy'ssolution I (ethanol and acetic acid), Carnoy's Solution II (ethanol,chloroform and acetic acid), methacarn (methanol, chloroform and aceticacid), Clark's fixative, Boonfix, and the like. A non-limiting list ofcommercially available fixatives includes, but is not limited to,MIRSKY'S FIXATIVE (available from National Diagnostics, Inc. of Atlanta,Ga.); GLYOFIX (available from Shandon Lipshaw, Inc. of Pittsburgh, Pa.);HISTOCHOICE (available from Amresco); HISTOFIX (available from TrendScientific, New Brighton, Minn.); KRYOFIX (available from Merck);MICROFIX (available from Energy Beam Sciences, Inc., East Granbury,Conn.); NEOFIX (available from Merck); NOTOX (available from Earth SafeIndustries, Inc., Belle Mead, N.J.); OMNIFIX II and OMNIFIX 2000(available from AnCon Genetics, Inc, Mellville, N.Y.); PREFER (availablefrom Anatech Ltd, Battle Creek, Mich.); PRESERVE (available from EnergyBeam Sciences, Inc., East Granbury, Conn.); SAFEFIX II (available fromThermo Fischer Scientific, Inc.); STATFIX (available from StatLabMedical Products, Inc. of Lewisville, Tex.); STF (Streck TissueFixative, available from Streck Laboratories, Omaha, Nebr.); UMFIX(available from Sakura Finetek USA, Inc., Torrance, Calif.); and FINEFIX(available from Milestone Medical of Shelton, Conn.). Commerciallyavailable stabilizers include, but are not limited to, RNALATER(available from Ambion, Inc., Austin Tex.); and RNEASY (available fromQiagen, Inc., Valencia, Calif.). Any other reagents known or hereafterdiscovered for use as fixatives and/or stabilizers are intended asuseful in the present invention.

In use, a biological sample, such as a tissue sample extracted from apatient for molecular or histology diagnostic testing, is placed withininternal cavity 54 of sample holder 50. In embodiments where sampleholder 50 is mated with closure 60, the tissue sample may be placedwithin sample holder 50 after closure 60 is removed from container 20,either with sample holder 50 still connected to closure 60 or afterfirst removing sample holder 50 from closure 60 and then reattachingthereto once the tissue sample has been placed in sample holder 50.Alternatively, if sample holder 50 is not mated with closure 60, sampleholder 50 containing the tissue sample therein can be directly placedinto container interior 36 once closure 60 has been removed. In thisembodiment, after sample holder 50 has been placed in container interior36, closure 60 should be used to enclose open end 24 of container 20.

It is noted that the Figures generally depict sample holder 50 as beingheld by receiving member 68 at a position generally adjacent the top endof container interior 36, however the precise location of sample holder50 within container interior 36 can be tailored depending on the volumeof solution within container interior 36. For example, it iscontemplated that receiving member 68 may be structured so as toposition sample holder 50 within container interior 36 at a locationsubstantially midway between first end 24 and second end 28. In thismanner, a smaller volume of solution within container interior 36 willstill ensure full contact of sample holder 50 (and any sample containedtherein) with any solution within container interior 36.

In embodiments including a one reagent scheme as discussed above, thetissue sample at this point is contained within sample holder 50 incontainer interior 36 in isolation from contact with any reagents. Whenit is desired to contact the tissue sample with the reagent, the userengages injection device 110 with port 76 and injects the reagentcontained within injection device 110 through port 76 and into containerinterior 36. Optionally, in order to allot for the increased volumewithin container interior 36 due to the introduction of the reagent frominjection device 110, vent 88 releases the necessary volume of air fromcontainer interior 36 immediately before or during the injectionprocess. After the tissue sample has been in contact with the reagentfor a desired time, closure 60 can be removed giving user access tosample holder 50 and the tissue sample therein for further diagnostictesting.

In embodiments including a two reagent scheme as discussed above, whensample holder 50 is placed within container interior 36, the tissuesample is placed in contact with the first reagent contained withincontainer interior 36, with such reagent flowing through fluid openings56 of sample holder 50, thereby contacting the tissue sample containedwithin internal cavity 54 thereof. The tissue sample can be maintainedin contact with the reagent within container interior 36 for a specifiedtime period after which time injection device 110 can be used to injecta second reagent into container interior 36 as discussed above. Again,to allot for the increased volume within container interior 36 due tothe introduction of the reagent from injection device 110, vent 88 mayrelease the necessary volume of media, preferably air, from containerinterior 36 immediately before or during the injection process. Inembodiments including a two reagent scheme, it may be advantageous thatthe total combined volume of reagent contained within container interior36 and injection device 110 be no more than the total volume ofcontainer interior 36 to ensure that all of the second reagent containedwithin injection device 110 enters container interior 36 without firsthaving to remove any reagent therefrom. After the tissue sample has hadsufficient contact with the second reagent, closure 60 can be removedgiving user access to the tissue sample for further diagnostic testing.

In embodiments where sample holder 50 is connected with closure 60,access to the tissue sample contained within sample holder 50 can beachieved by removing closure 60 from container 20 and inverting it,placing the outer surface on a counter, thereby providing sample holder50 exposed. Any fluid that is contained within sample holder 50 can dripdownward within the bottom or internal surface of closure 60 and becaught by the rim surrounding closure 60, thereby preventing any leakageor spillage onto the counter surface. Sample holder 50 may be openablewhile sample holder 50 is connected with closure 60, thereby providingsimple access to the tissue sample contained therein, and providing aproper support for maintaining sample holder 50 in place without havingto physically contact any portion of sample holder 50 to hold it inplace while accessing the sample, thus preventing any potential forcontamination of the sample based on contact by the user.

Thereafter, container 20 may be washed and reused, or more preferably,will be discarded to prevent cross-contamination with other samples.Injection device 110 may be refilled with, preferably, the same solutionand then reused or it may be discarded.

While embodiments of the present invention are satisfied in manydifferent forms, there is shown in the figures and described herein indetail, specific embodiments of the invention, with the understandingthat the present disclosure is to be considered as exemplary of theprinciples of the invention and is not intended to limit the inventionto the embodiments illustrated. Various other embodiments will beapparent to, and readily made by those skilled in the art, withoutdeparting from the scope and spirit of the invention. For example, whilethe disclosure and drawings depict a system including one injectiondevice to inject a reagent into the container, it is contemplated thatadditional injection devices may be provided, such that the system mayinclude any number of injection devices with varying reagents. The scopeof the invention will be measured by the appended claims and theirequivalents.

1. A system for storing a biological sample, comprising: a containerhaving a closed end, an open end, and a sidewall extending therebetween,defining a container interior; a removable closure for enclosing theopen end of the container; a sample holder for housing a biologicalsample, insertable within the container interior; a port disposed withinone of the container and the removable closure; and an injection deviceengageable with the port to allow a fluid to pass from within theinjection device into the container interior.
 2. The system of claim 1,wherein sample holder is detachably connected to the removable closure.3. The system of claim 2, wherein the port is disposed within theclosure adjacent the sample holder.
 4. The system of claim 1, furthercomprising a platform attached to the removable closure and adapted forreceiving the sample holder.
 5. The system of claim 1, wherein thesample holder is positioned in fluid-access with the port such thatfluid introduced through the port passes directly through the sampleholder.
 6. The system of claim 1, wherein the sample holder is ahistology cassette.
 7. The system of claim 1, further comprising a ventadapted to discharge a volume of air from the container interior.
 8. Thesystem of claim 7, wherein the vent is disposed within the removableclosure.
 9. The system of claim 7, wherein the vent transitions from afirst closed position to a second open position when the volumetricpressure within the container interior reaches a threshold value. 10.The system of claim 1, wherein the removable closure and the open end ofthe container are threadably matable.
 11. The system of claim 1, whereinthe injection device is a syringe.
 12. The system of claim 1, whereinthe injection device engages the port and, upon engagement, the porttransitions from a first closed position to a second open position. 13.The system of claim 1, further comprising a first fluid disposed withinthe container interior.
 14. The system of claim 13, further comprising asecond fluid disposed within the injection device, the second fluiddifferent than the first fluid.
 15. The system of claim 1, wherein theport comprises a luer-activated valve.
 16. A container for storing abiological sample, comprising: a housing having a closed end, an openend, and a sidewall extending therebetween, defining a containerinterior; a removable closure for enclosing the open end of the housing;a sample holder for housing a biological sample, detachably connected tothe removable closure and insertable within the container interior; anda port disposed within the removable closure adjacent the sample holder,the port adapted to allow a fluid to pass therethrough from a locationexterior to the container to the container interior.
 17. The containerof claim 16, wherein the sample holder is a histology cassette.
 18. Thecontainer of claim 16, further comprising a vent disposed within theremovable closure and adapted to discharge a volume of air from thecontainer interior.
 19. The container of claim 16, further comprising afirst fluid disposed within the container interior.
 20. The container ofclaim 19, wherein the port is adapted to receive an injection device atleast partially therein, for transferring a second fluid into thecontainer interior, the second fluid different than the first fluid. 21.A method of storing a biological sample, comprising the steps of:providing a container comprising: a housing having a closed end, an openend, and a sidewall extending therebetween defining a container interiorhaving a first fluid disposed therein, a removable closure for enclosingthe open end of the housing, a sample holder housing a biologicalsample, the sample holder detachably connected to the removable closureand disposed within first fluid of the container interior, and a portdisposed within the removable closure adjacent the sample holder;engaging an injection device containing a second fluid with the port,the second fluid different than the first fluid; and injecting thesecond fluid from the injection device through the port into thecontainer interior.